著者
坂野 好幸 長畑 直和 藤本 大三郎
出版者
The Japanese Society of Applied Glycoscience
雑誌
澱粉科学 (ISSN:00215406)
巻号頁・発行日
vol.38, no.2, pp.187-192, 1991-06-30 (Released:2011-07-01)
参考文献数
22

Pseudomonas amyloderamosa isoamylase and Klebsiella pneumoniae pullulanase were crystalline preparations obtained from Hayashibara Biochemical Laboratory, Inc., Okayama, Japan. Bacillus acidopullulyticus pullulanase was purified from Promozyme 200 L (Novo Nordisk Bioindustry, Ltd., Copenhagen, Denmark) by the method of Kusano et al. (Agric. Biol. Chem., 52, 2293). These three enzyme preparations showed a single band on PAGE. Optimum pH of Pseudomonas isoamylase for amylopectin was 3.5 with a shoulder near pH 5. The optimum pH for Br-CDs shifted from 3.5 for amylopectin to 4.5-4.7 (Br-α- and -β-CDs) and 4.0 (G3-, G4-γ-CDs). The pH curve for Br-γ-CDs had a shoulder near pH 5.0. Optimum pHs of Klebsiella and Bacillus pullulanases for pullulan and Br-γ-CDs were 5.5 and 5. 0, but those for Br-α-CDs were 6.0 and 4. 0, respectively. Kinetic parameters of these enzymes for Br-CDs indicated that (1) all of them cleaved more easily the a(1→6) linkages of G3-G5-CDs than those of G2-CDs, (2) they split more easily the a(1→6) linkages of Br-r-CDs than those of the other Br-CDs, (3) Pseudomonas isoamylase hydrolyzed readily the a(1→6) linkage of G2-7-CD and (4) Br-r-CDs were better substrates for kinetical analysis of debranching amylase than amylopectin and pullulan.